RT Journal Article SR Electronic T1 An improved Nicotiana benthamiana strain for aphid and whitefly research JF bioRxiv FD Cold Spring Harbor Laboratory SP 2020.08.04.237180 DO 10.1101/2020.08.04.237180 A1 Honglin Feng A1 Lucia Acosta-Gamboa A1 Lars H. Kruse A1 Alba Ruth Nava Fereira A1 Sara Shakir A1 Hongxing Xu A1 Garry Sunter A1 Michael A. Gore A1 Gaurav D. Moghe A1 Georg Jander YR 2020 UL http://biorxiv.org/content/early/2020/08/05/2020.08.04.237180.abstract AB Nicotiana benthamiana is used extensively as a platform for transient gene expression and as a model system for studying plant-virus interactions. However, many tobacco-feeding insects, e.g. Myzus persicae (green peach aphids) and Bemisia tabaci (whiteflies), grow poorly on N. benthamiana, limiting its utility for research on plant-insect interactions. Using CRISPR/Cas9, we generated knockout mutations in two N. benthamiana acylsugar acyltransferase genes, ASAT1 and ASAT2, which contribute to the biosynthesis of insect-deterrent acylsucroses. Whereas ASAT1 mutations reduced the abundance of two predominant acylsucroses, ASAT2 mutations caused almost complete depletion of foliar acylsucroses. Both M. persicae and B. tabaci survived and reproduced significantly better on asat2 mutant plants than on wildtype N. benthamiana. Furthermore, ASAT1 and ASAT2 mutations reduced the water content and increased the temperature of leaves, indicating that foliar acylsucroses can protect against desiccation. Improved aphid and whitefly performance on ASAT2 mutants will make it possible to use the efficient transient overexpression and gene expression silencing systems that are available for N. benthamiana to study plant-insect interactions. Additionally, the absence of acylsugars in ASAT2 mutant lines will simplify transient expression assays for the functional analysis of acylsugar biosynthesis genes from other Solanaceae.Competing Interest StatementThe authors have declared no competing interest.